富贵竹过氧化物酶全长基因的克隆与表达分析

Molecular Cloning and Expression Patterns of Full-length Peroxidases Gene from Dracaena sanderiana

[目的]克隆富贵竹全长POD基因,检测富贵竹感染斯氏泛菌(Pantoea stewartii)后该基因的表达。[方法]用PCR克隆富贵竹(Dracaena sanderiana)POD部分基因,用hiTAIL-PCR扩增该基因左右两侧序列。通过RT-PCR和qPCR确定POD基因在病原物侵染时的表达。[结果]获得一个长度为3419 bp的序列,登录号为MZ450796。经基因结构预测,该序列包含1个转录起始位点、加尾信号和4个外显子。进化树分析表明,该基因编码的氨基酸序列与芦笋POD的氨基酸序列一致性最高。接种P.stewartii后,POD基因的表达上调,3 d达最大值。[结论]克隆了富贵竹全长POD基因,确定了POD基因在富贵竹叶片中表达和结构预测的正确性;POD基因被P.stewartii诱导上调表达。

[Objective]Peroxidase(POD)plays an important role in the resistance of plant disease.The full-length POD gene was cloned and the expression of the gene was determined after Dracaena sanderiana was infected by Pantoea stewartii.[Method]The partial POD gene of D.sanderiana was cloned by PCR.HiTAIL-PCR was used to amplify the left and right parts of the POD gene.The expression of the gene was determined by RT-PCR and qPCR.[Result]A 3419 bp sequence was obtained,the accession number was MZ450796.The sequence contained a transcription start site,a polyadenylation signal and four exons after the gene structure prediction.Phylogenetic tree analysis showed that the amino acid sequence encoded by this gene had the highest identity with that of Asparagus officinalis POD.The expression of the POD gene of D.sanderiana was up-regulated by P.stewartia and reached the maximum after 3 days.[Conclusion]The full-length POD gene is obtained.The POD gene expresses in the leaf of D.sanderiana and the predicted gene structure is correct.The expression of the POD gene of D.sanderiana was induced by P.stewartia.